CN107141323A - Reduction/pH dual responsiveness adriamycin prodrugs and preparation method and application - Google Patents
Reduction/pH dual responsiveness adriamycin prodrugs and preparation method and application Download PDFInfo
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- A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6905—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a colloid or an emulsion
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- C08G65/00—Macromolecular compounds obtained by reactions forming an ether link in the main chain of the macromolecule
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- C08G65/32—Polymers modified by chemical after-treatment
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- C08G65/334—Polymers modified by chemical after-treatment with organic compounds containing sulfur
Abstract
The invention discloses a kind of reduction/pH dual responsiveness adriamycin prodrugs and preparation method and application.Specific preparation method is the sulphur diine butyl dipropionate of micromolecular compound 3,3 ' two blocked using alkynyl（B‑ss‑B）The acetal radical polyethylene glycol of two azidoethyl of compound two blocked with azido（N3‑a‑PEG‑a‑N3）Chemically reacted, prepare alkynyl end-blocking has reduction/pH dual responsiveness polymer poly (SSalt‑A)n；Then doxorubicin derivative is connected to the two ends of polymer segment, reduction/pH dual responsiveness adriamycin prodrugs DOX is obtainedhyd‑poly(SS‑alt‑A)n‑hyd‑DOX.This kind of water-soluble adriamycin prodrug with reduction/pH dual responsiveness has the characteristic of good biocompatibility and controllable in drug release, thus may be used as stimulating sensitiveness anti-tumor predrug.
Description
Technical field
The invention belongs to field of biomedical polymer materials, and in particular to a kind of reduction/pH based on polyethylene glycol is double
Weight response adriamycin prodrug, its preparation method and its application.
Background technology
Cancer is that body causes the result of normal cell canceration in many factors, multiple stages and multiple mutation, in doctor
Malignant tumour is also referred to as on.The lethality of cancer is big, easily transfer, by destroying the various functions of tissue and organ, finally
People are caused to be died because of dying of exhaustion for organ dysfunction.According to statistics, with the growth of population, the incidence of disease of cancer is also constantly increasing
It is long, the health of the mankind is seriously endangered, therefore, cancer treatment method enjoys the attention of domestic and international medical investigators.
In in the past few decades, with the progress of medical technology, the treatment method of various cancers is developed rapidly,
Including operative treatment, chemotherapy, radiotherapy, immunization therapy, hormone therapy, targeted therapy etc..But, make in chemotherapy process
Small-molecule drug（Such as：Taxol, adriamycin, camptothecine etc.）, to normal cell while tumour cell effect is killed
Also have very strong damaging action, therefore, can cause it is serious vomit, the side effect such as dizzy, body function declines.Meanwhile, small point
Sub- medicine is easily recognized and discharged by body during blood circulation, adds environment special near tumor tissues, such as crisscross multiple
Miscellaneous blood vessel network structure, fine and close interstitial structure and higher interstitial fluid pressure etc. so that medicine is difficult that arrival tumour is thin
Intracellular.The curative effect of antineoplastic clinically is have impact on just because of these restrictive conditions.
By prodrug, above mentioned problem can be efficiently solved.Prodrug (prodrug), also referred to as prodrug, premedicant
Thing, refers to there is the compound of pharmacological action after organism conversion.Polymeric prodrugs refer to active medicine with rising
The polymer support of translocation makes medicine without activity by Covalent bonding together.But, when prodrug enter body circulation and
In metabolic process, active medicine can be discharged by hydrolysis removal carrier, play pharmacological action.This mode can have
Effect ground improves the utilization rate of medicine, strengthens targeting, reduces the toxic side effect of medicine.
In polymeric prodrugs structure, the selection for polymeric base material is most important, and biocompatibility is material
Most basic requirement for pharmaceutical carrier.Polyethylene glycol（PEG）With non-toxic, good biocompatibility and biodegradable
Property, used by food and medicine Surveillance Authority of the U.S. (FDA) approval.By carrying out end-functionalization modification to PEG, then with dredging
Aqueous pharmaceutical is bonded, and can prepare polymeric prodrugs, assigns drug molecule some advantageous properties.Although polymeric prodrugs are in cancer
There is very big advantage in treatment, still, how more drug is delivered in tumour cell, is still to receive much concern always
Problem.Medicine, which reaches lesion site, to be needed to overcome various barriers, is considered as during blood circulation how keeping transporting carrier
Stability, extend circulation time；How efficiently to enter intracellular, more accumulate at specific position；How to medicine
Controlled release etc..If not solving these problems, therapeutic effect of the medicine to cancer will be greatly reduced.
In the prior art, some are had on sensitive and sensitivity to acid prodrug the report of reduction.But, before antitumor
Medicine, should have good biocompatibility and biodegradability, also, as anti-tumor predrug, should also have following
Feature：Stable polymer micelle can be formed in aqueous, and hydrophily shell plays stable micella, raising micella blood and followed
The effect of ring time；When prodrug micelle is circulated in vivo, the performance with anticoagulation and anti-protein adsorption；When carrier micelle is reached
When tumour or pathological tissues, the characteristics of can utilizing the low pH conditions of local organization and high glutathione concentrations destroys micella,
Quick release goes out cancer therapy drug.Accordingly, it would be desirable to seek more drug effects substantially and in tumour cell microenvironment have reduction and
Sensitivity to acid anti-tumor predrug.
The content of the invention
It is an object of the invention to provide a kind of reduction based on polyethylene glycol/pH dual responsiveness adriamycin prodrugs and its system
Preparation Method；The adriamycin prodrug has good biocompatibility and suppresses the ability of tumor cell proliferation.
Specifically technical scheme is the present invention：A kind of reduction/pH dual responsiveness adriamycin prodrugs, by the following chemical structure
Formula is expressed：
M is that 3~113, n is 3~15.
In above-mentioned technical proposal, adriamycin prodrug has the group of sensitivity to acid and the group of reduction-sensitive；The tool
The hydrophobicity adriamycin part that the adriamycin prodrug for having dual responsiveness contains in structure, the kernel for forming micella, PEG
Hydrophilic parts are wrapped in the shell that micella is formed outside kernel, have good effect to the stability of micella.Reduction and
Under the conditions of pH, acid sensitive group and reduction sensitive group are broken, and micella is destroyed, and is gathered in so as to rapidly discharge
Hydrophobic anticancer drug inside micella.
It is preferred that technical scheme in, the number-average molecular weights of reduction/pH dual responsiveness adriamycin prodrugs is 4000~80000
g·mol-1。
The present invention is using the alternate copolymer poly (SS- containing acid sensitive group and reduction sensitive group in structurealt- A) as base stock, reacted after mantoquita and ligand catalysis chemical reaction activation with anticancer drugs, doxorubicin, prepare base
In reduction/pH dual responsiveness adriamycin prodrugs DOX- of polyethylene glycolhyd-poly(SS-alt-A)-hyd-DOX。
The preparation method of above-mentioned reduction/pH dual responsiveness adriamycin prodrugs, comprises the following steps：
(1) with 6- nitrine hexanoyl hydrazines and doxorubicin hydrochloride（DOX×HCl）As raw material, absolute methanol is solvent, glacial acetic acid
For catalyst, doxorubicin derivative is obtained by chemically reacting；
Wherein, the mol ratio of 6- nitrine hexanoyl hydrazine and doxorubicin hydrochloride is 1：(2~6)；
The chemical structural formula of the 6- nitrine hexanoyl hydrazine is：
The chemical structural formula of the doxorubicin derivative is：
(2) under the conditions of inert gas atmosphere, in the presence of copper salt catalyst and part, with 3,3 '-two sulphur diine butyl dipropyls
Acid esters and the acetal radical polyethylene glycol of two azidoethyl two are raw material, using anhydrous DMF as solvent, pass through reaction
Prepare the polyethylene glycol alternate copolymer of two end alkynyl radicals end-blocking；
Wherein, the acetal radical polyethylene glycol of two azidoethyl two, 3,3 '-two sulphur diine butyl dipropionates and copper salt catalyst are rubbed
You are than being 1: (1.1~1.5):（0.5~1.5）；Copper salt catalyst and the mol ratio of part are 1: (1~2)；
The chemical structural formula of 3,3 '-two sulphur diine butyl dipropionate is：
The structural formula of the acetal radical polyethylene glycol of two azidoethyl two is：
Above-mentioned m is 3~113；
The structural formula of the polyethylene glycol alternate copolymer of two end alkynyl radical end-blocking is：
Above-mentioned m is that 3~113, n is 3~15.
(3) using anhydrous DMF as solvent, in the presence of copper salt catalyst and part, step is used
(2) the polyethylene glycol alternate copolymer of the two end alkynyl radicals end-blocking obtained reacts with the doxorubicin derivative that step (1) is obtained, system
It is standby to obtain the reduction/pH dual responsiveness adriamycin prodrugs；
The polyethylene glycol alternate copolymer of the two end alkynyl radicals end-blocking, the mol ratio of doxorubicin derivative are 1：(2~5).
In above-mentioned technical proposal, doxorubicin derivative is prepared（N3-hyd-DOX×HCl）, mainly prepared by three steps
Form：
The first step, prepares 6- nitrine methyl caproates.With 6- bromocaproic acids methyl esters and sodium azide（NaN3）Reaction, using DMF as solvent,
Azido reaction is carried out, 6- nitrine methyl caproates are obtained.Wherein, 6- bromocaproic acids methyl esters, the mol ratio of sodium azide are 1：(1.5
~5)；
Second step, prepares 6- nitrine hexanoyl hydrazines.The hydration of the 6- nitrine methyl caproate synthesized using the first step and mass fraction as 85%
Hydrazine reaction, with tetrahydrofuran（THF）As solvent, by amidation process, 6- nitrine hexanoyl hydrazines are obtained.Wherein, 6- nitrine oneself
The mol ratio of sour methyl esters and hydrazine hydrate is 1：(10~30)；
3rd step, prepares doxorubicin derivative（N3-hyd-DOX×HCl）.The 6- nitrine hexanoyl hydrazines that are synthesized using second step and Ah
Mycin hydrochloride（DOX×HCl）As raw material, using absolute methanol as solvent, glacial acetic acid is catalyst, obtains adriamycin and derives
Thing.Wherein, the mol ratio of 6- nitrine hexanoyl hydrazine and doxorubicin hydrochloride is 1：(2~6)；
Preparing the polyethylene glycol alternate copolymer of two end alkynyl radicals end-blocking includes the following steps：
Prepare alkynyl the is blocked and sulphur diine butyl dipropionate of micromolecular compound 3,3 '-two containing disulfide bond（B-ss-
B）.Be raw material using 3,3 '-dithiodipropionic acid and 3- butyne-1-ols, with 1- ethyls-（3- dimethylaminopropyls）Carbon two is sub-
Amine hydrochlorate is activator, and DMAP is catalyst, and dichloromethane is solvent, by esterification, obtain 3,3 '-
Two sulphur diine butyl dipropionates.Wherein, 3, the mol ratio of 3 '-dithiodipropionic acid and 3- butyne-1-ols is 1：(1.5~3)；
Prepare the acetal radical polyethylene glycol of two azidoethyl two of azido end-blocking（N3-a-PEG-a-N3）, carried out by two steps：
The first step, synthesizes the acetal radical polyethylene glycol of Dichloroethyl two（Cl-a-PEG-a-Cl）.Using polyethylene glycol（HO-PEG-
OH）With 2- chloroethyl vinyl ethers（CEVE）In the presence of raw material, acid catalyst, reaction obtains the polyethylene glycol that end is chloro
Product Cl-a-PEG-a-Cl.Wherein, the molecular weight of polyethylene glycol is：400~5000 g·mol-1, polyethylene glycol and 2- chloroethyls
The mol ratio of vinyl ethers is 1：(2~6)；Acid catalyst is in p-methyl benzenesulfonic acid, p-methyl benzenesulfonic acid pyridine, trifluoroacetic acid
One kind；
Second step, synthesizes the acetal radical polyethylene glycol of two azidoethyl two（N3-a-PEG-a-N3）.The Cl- that the first step is obtaineda-
PEG-a- Cl and reaction of sodium azide, using dichloromethane as solvent, obtain the acetal radical polyethylene glycol of two azidoethyl two（N3-a-
PEG-a-N3）.Wherein, polyethylene glycol of the end containing chloro（Cl-a-PEG-a-Cl）Mol ratio with sodium azide is 1：
(2~6)；
Prepare the polymer poly (SS- of alkynyl end-blockingalt-A).Under nitrogen protection, in the presence of copper salt catalyst and part
Under, utilize the 3 of above-mentioned synthesis, 3 '-two sulphur diine butyl dipropionates（B-ss-B）With the poly- second two of the acetal radical of two azidoethyl two
Alcohol（N3-a-PEG-a-N3）Chemically react, prepare the polymer poly (SS- of alkynyl end-blockingalt-A).Wherein, two nitrine second
The acetal radical polyethylene glycol of base two（N3-a-PEG-a-N3）With 3,3 '-two sulphur diine butyl dipropionates（B-ss-B）Mol ratio be
1：(1.1~1.5)；
Prepare the adriamycin prodrug DOX- of double-responsehyd-poly(SS-alt-A)-hyd- DOX includes the following steps：
The polyethylene glycol alternate copolymer of the two end alkynyl radicals end-blocking synthesized using the doxorubicin derivative of desalting processing with upper step
poly(SS-alt- A) chemically react, obtain reduction/pH dual responsiveness adriamycin prodrugs DOX-hyd-poly(SS-alt-A)-hyd-DOX.Wherein, the polyethylene glycol alternate copolymer of two end alkynyl radicals end-blocking and the mol ratio of doxorubicin derivative are
1：(2~5).
In above-mentioned technical proposal, in step (1), reaction temperature is 60 DEG C~80 DEG C, and the reaction time is the h of 12 h~24；Step
Suddenly in (2), inert gas is nitrogen；Reaction temperature is 25 DEG C~50 DEG C, and the reaction time is the h of 24 h~72；In step (3), instead
It is 25 DEG C~50 DEG C to answer temperature, and the reaction time is the h of 24 h~48.
In above-mentioned technical proposal, in step (2) and step (3), copper salt catalyst is selected from Salzburg vitriol, stannous chloride
Or cuprous bromide；Part is selected from：Sodium ascorbate, bipyridyl, five methyl diethylentriamine, tetramethylethylenediamine or hexamethyl
One kind in trien.
The present invention in the presence of copper salt catalyst and part, by the restriction of raw material and parameter, is prepared first
Polyethylene glycol alternate copolymer containing acid sensitive group and reduction sensitive group, solving existing reaction system can not prepare
Go out simultaneously containing acid sensitive group and the problem of reduce the alternate copolymer of sensitive group, be it is a kind of it is new, simple, efficient,
Quick synthetic method.
Further technical scheme, after the completion of step (1)~(3), carries out purification processes, the purifying to product respectively
Process comprises the following steps：
(i) doxorubicin derivative N3-hyd- DOX × HCl purifying：After reaction terminates, reaction solution is filtered to remove precipitation with cotton
Thing, removes and is precipitated three times in ice ether after solvent；Sediment is obtained using centrifugal process, places it in vacuum drying chamber and does
The dry product N for obtaining red powder3-hyd-DOX×HCl；
(ii) the polyethylene glycol alternate copolymer poly (SS- of two end alkynyl radicals end-blockingalt- A) purifying：After reaction terminates, use
DMF dialyses after 48 h, is placed in 60 DEG C of oil baths, DMF is removed under reduced pressure；Then with 150 mL CH2Cl2Again dissolve, use saturation
The NaCl aqueous solution is extracted, and removes mantoquita；Organic layer anhydrous Na2SO4Dry after 4 h, rotary evaporation removing solvent, then
Precipitated 3 times in diethyl ether solution, sediment is collected and is dried in vacuo, sticky product poly (SS- are finally givenalt-A)；
(III) reduction/pH dual responsiveness adriamycin prodrugs DOX-hyd-poly(SS-alt-A)-hyd- DOX purifying：Reaction
After end, using deionized water 72 h of dialysis, the red, transparent liquid freezing obtained by bag filter is dried, peony is obtained
Solid product DOX-hyd-poly(SS-alt-A)-hyd-DOX。
In above-mentioned technical proposal, in step (i), solvent is removed using the method for rotary evaporation；When being dialysed in step (ii)
Molecular cut off is used for 1000~14000 Da bag filter；In step (III), molecular cut off is used during dialysis for 3500
~14000 Da bag filter.
Specific purification comprises the following steps：
(i) doxorubicin derivative is prepared（N3-hyd-DOX×HCl）Purifying：
The first step, prepares the purifying of 6- nitrine methyl caproates：After reaction terminates, filtered and reacted with the short column of neutral alundum (Al2O3)
Liquid, and with oil pump removal of solvent under reduced pressure DMF.Then, crude product is dissolved with substantial amounts of dichloromethane, is extracted with 30 mL secondary water
Take three times, organic phase is collected and the h of anhydrous sodium sulfate drying 4 is used.Finally, filtered with cotton, rotary evaporation removes solvent, will
Product, which is placed in vacuum drying oven, dries 24 h, obtains colourless product liquid 6- nitrine methyl caproates；
Second step, prepares the purifying of 6- nitrine hexanoyl hydrazines：After reaction terminates, rotary evaporation uses CH instead after removing THF2Cl2Dissolving.
In separatory funnel, crude product is extracted using saturated sodium-chloride water solution.By organic phase anhydrous Na2SO44 h are dried, are removed molten
It is dried in vacuo after agent, is finally collected into product 6- nitrine hexanoyl hydrazines；
3rd step, prepares the purifying of doxorubicin derivative：After reaction terminates, the Na in reaction solution is filtered to remove with cotton2SO4,
Filtrate rotary evaporation is removed after solvent, precipitated three times in ice ether.Finally, following sediment is obtained using centrifugal process,
Place it in and 24 h are dried in vacuum drying chamber, obtain red powder product, as N3-hyd-DOX×HCl；
(ii) 3,3 '-two sulphur diine butyl dipropionates of alkynyl end-blocking are prepared（B-ss-B）Purifying：After reaction terminates, use
Filter paper is filtered, except the salt generated in dereaction, then with 150 mL CH2Cl2Dissolving, with 1 M HCl solutions and saturated sodium-chloride
（NaCl）The mixed solution of the aqueous solution is extracted 2 times, and each water layer continues the CH with 30 mL2Cl2Extraction 3 times.Organic layer is collected
Afterwards, anhydrous Na is used2SO44 h are dried, the crude product after solvent is removed and continues to carry out separating-purifying using column chromatography.Collect final
Product, is dried in vacuo 24 h, that is, obtains 3,3 '-two sulphur diine butyl dipropionates（B-ss-B）；
(III) prepares the acetal radical polyethylene glycol of two azidoethyl two of azido end-blocking（N3-a-PEG-a-N3）Purifying：
The first step, prepares the synthesis acetal radical polyethylene glycol of Dichloroethyl two（Cl-a-PEG-a-Cl）Purifying：After reaction terminates,
First use CH2Cl2Dilute reaction solution, with 10 mL phosphate buffer solution（PBS, pH 10.0) and the saturation NaCl aqueous solution mixing
Solution is extracted, and obtained water layer continues to use CH2Cl2Extraction three times.Organic layer anhydrous Na2SO4Dry after 4 h, rotary evaporation is removed
A large amount of solvents are removed, when remaining about 3 mL crude product in solution, it is precipitated three times in n-hexane, sediment is collected and is dried in vacuo,
Obtain product Cl-a-PEG-a-Cl；
Second step, prepares the synthesis acetal radical polyethylene glycol of two azidoethyl two（N3-a-PEG-a-N3）Purifying：Reaction terminates
Afterwards, neutral Al is used2O3Short column be filtered to remove unreacted NaN3, solvent DMF is removed under 60 °C of oil bath reduced pressures.Then,
Use CH2Cl2Crude product is dissolved, with pH 10.0 PB cushioning liquid purification by liquid extraction products, each water layer continues with 20 mL's
CH2Cl2Extraction, most organic phase is collected and uses anhydrous Na at last2SO4Dry, rotary evaporation, which removes solvent and is dried in vacuo, obtains palm fibre
The product N of color3-a-PEG-a-N3；
The polyethylene glycol alternate copolymer poly (SS- of (IV) two end alkynyl radical end-blockingalt- A) purifying：After reaction terminates, use
DMF dialyses after 48 h, is placed in 60 DEG C of oil baths, DMF is removed under reduced pressure.Then with 150 mL CH2Cl2Again dissolve, use saturation
The NaCl aqueous solution is extracted, and removes mantoquita.Organic layer anhydrous Na2SO4Dry after 4 h, rotary evaporation removing solvent, then
Precipitated 3 times in ether, sediment is collected and is dried in vacuo, sticky product poly (SS- are finally givenalt-A)；
(V) prepares the adriamycin prodrug DOX- of double-responsehyd-poly(SS-alt-A)-hyd- DOX purifying：Reaction knot
Shu Hou, using deionized water 72 h of dialysis, the red, transparent liquid freezing obtained by bag filter is dried, Red viscous is obtained
Solid product DOX-hyd-poly(SS-alt-A)-hyd-DOX。
Reduction disclosed by the invention/pH dual responsiveness adriamycin prodrugs DOX-hyd-poly(SS-alt-A)-hyd-DOX
Prodrug micelle, the core of hydrophobic drug adriamycin formation micella can be self-assembly of in aqueous；Hydrophily PEG chain section shape
Into the shell of micella, play a part of stable micella.And contain reduction response between hydrophobic cores and hydrophily PEG chain section
The acetal and acylhydrazone key group of disulfide bond and acid-sensitive, under the conditions of reduction and pH, sensitive group is broken, and micella is broken
It is bad, so as to rapidly discharge the hydrophobic anticancer drug being gathered in inside micella.Therefore, the present invention is claimed above-mentioned simultaneously
Reduction/application of the pH dual responsiveness adriamycin prodrugs in stimulating responsive anticancer nano medicine is prepared.
Due to the implementation of such scheme, the present invention compared with prior art, with advantages below：
1st, the present invention uses the good polyethylene glycol of biocompatibility and hydrophobic anticancer drug adriamycin respectively as hydrophily
Segment and hydrophobic end, to be used as sensitiveness linker with the group that can be hydrolyzed under acid condition under the reducing conditions
Group, is prepared for the reduction with good biocompatibility/pH dual responsiveness adriamycin prodrugs；It is preceding compared with active compound adriamycin
Medical instrument has good water solubility and bin stability, easily decomposes in tumour cell environment, accelerates insoluble drug release, greatly increases
Its application value.
2nd, reduction/pH dual responsiveness adriamycin prodrugs that the present invention is obtained can be self-assembly of stable polymerization in water
Thing prodrug micelle.Hydrophobic adriamycin moiety aggregation formation micelle inner core, hydrophily PEG chain section is played surely as micella shell
Determine micella, improve the effect of micella blood circulation time.Hydrophilic segment and hydrophobic part pass through cleavable in acid condition
Key is learned to be connected, at the same inside hydrophilic PEG chain segment containing under the reducing conditions with the group that can be hydrolyzed under acid condition；When
When prodrug micelle reaches tumour or pathological tissues, because the pH value of these tissue microenvironments is reduced, acid sensitive group can be made
Fracture, while in the environment of high glutathione, reduction sensitive group is also broken, and causes micellar structure to be destroyed, from
And quick release goes out cancer therapy drug, the utilization rate and targeting of medicine are added, in terms of the treatment of cancer, is answered with potential
With value.
3rd, clearly, synthesis condition is not harsh, has for reduction/pH dual responsiveness adriamycins pro-drugs that the present invention is provided
Following distinguishing feature：(1) raw materials used and reagent is readily available；(2) reaction condition is simple；(3) yield is high；(4) it is three-dimensional
Selectivity is good；(5) product separation is easy；(6) product stability is good；It is easy to prepare, and purification is convenient, is adapted to industrialized production.
Brief description of the drawings
Fig. 1 is doxorubicin derivative in embodiment one（N3-hyd-DOX×HCl）Hydrogen nuclear magnetic resonance spectrogram, solvent is deuterium
For dimethyl sulfoxide；
Fig. 2 is the hydrogen nuclear magnetic resonance spectrogram of 3,3 '-two sulphur diine butyl dipropionates in embodiment two, and solvent is deuterochloroform；
Fig. 3 is Cl- in embodiment threea-PEG21-a- Cl and N3-a-PEG21-a-N3Hydrogen nuclear magnetic resonance spectrogram, solvent is deuterated
Chloroform, wherein, subscript 21 represents-CH2CH2O- number of repeat unit；
Fig. 4 is polymer poly (SS- in example IValt-A)3.6Hydrogen nuclear magnetic resonance spectrogram, solvent is deuterochloroform, its
In, subscript 3.6 represents the degree of polymerization of alternate copolymer；
Fig. 5 is DOX- in embodiment fivehyd-poly(SS-alt-A)3.6-hyd- DOX hydrogen nuclear magnetic resonance spectrogram, solvent is deuterium
For chloroform；
Fig. 6 is HO-PEG in embodiment one~five21-OH, Cl-a-PEG21-a-Cl, N3-a-PEG21-a-N3, poly(SS-alt-A)3.6, N3-hyd- DOX × HCl and DOX-hyd-poly(SS-alt-A)3.6-hyd- DOX infrared spectrogram；
Fig. 7 is DOX- in embodiment fivehyd-poly(SS-alt-A)3.6-hyd- DOX the self assembly in the cushioning liquid of pH 7.4
The dynamic light scattering curve and transmission electron microscope photo figure of the micella of formation；
Fig. 8 is DOX- in embodiment fivehyd-poly(SS-alt-A)3.6-hydThe polymeric prodrugs micella and naked medicine of-DOX formation
Drug release patterns figure of the adriamycin in the cushioning liquid of different pH value；
Fig. 9 is DOX-hyd-poly(SS-alt-A)3.6-hydThe suppression tumor cell proliferation of-DOX prodrugs and naked medicine adriamycin
Can test chart.
Embodiment
With reference to embodiment and accompanying drawing, the invention will be further described：
Embodiment one：Doxorubicin derivative（N3-hyd-DOX×HCl）Synthesis
Doxorubicin derivative（N3-hyd-DOX×HCl）Mainly it is prepared from by three steps.The first step, prepare 6- nitrine oneself
Sour methyl esters；Second step, prepares 6- nitrine hexanoyl hydrazines；3rd step, to doxorubicin hydrochloride（DOX×HCl）Progress modify obtain Ah
Adm derivative（N3-hyd-DOX×HCl）, specific synthetic method is as follows：
Synthesized micromolecule compound 6- nitrine methyl caproates.The device for being positioned over 120 °C of baking ovens in advance is taken out, guarantor is put into dry
In device, treat that it is cooled to room temperature, taking-up is used.6- bromocaproic acid methyl esters is weighed respectively（3.54 g, 16.93 mmol）And Azide
Sodium（NaN3）（2.75 g, 42.33 mmol）, and be dissolved in 20 mL DMF in 50 mL round-bottomed flask, in 60 °C of condensations
React and terminate after 12 h under water counterflow condition.Use neutral alundum (Al2O3)（Al2O3）Short column filtering reacting liquid, remove unreacted
NaN3, and with oil pump removal of solvent under reduced pressure DMF.Then, substantial amounts of dichloromethane is used（CH2Cl2）Crude product is dissolved, 30 mL are used
Secondary water extract three times, organic phase is collected and anhydrous sodium sulfate is used（Na2SO4）Dry 4 h.Finally, filtered, revolved with cotton
Turn evaporation of solvent, product is placed in vacuum drying oven and dries 24 h, colourless product liquid 6- nitrine methyl caproates are obtained
（2.47 g, yield：53.2%）；
Obtained 6- nitrine methyl caproates are added in round-bottomed flask（0.76 g, 4.44 mmol）It is 85% with mass fraction
Hydrazine hydrate（5.60 g, 111.1 mmol）, using tetrahydrofuran（THF）It is used as solvent.Equally, in 80 °C of oil baths and equipped with snake
In the device of shape condenser pipe, the h of back flow reaction 12.After reaction terminates, rotary evaporation removes THF, uses CH2Cl2Dissolving.In a point liquid leakage
In bucket, crude product is extracted using saturated sodium-chloride water solution.By organic phase anhydrous Na2SO44 h are dried, vacuum after solvent is removed
Dry, be finally collected into product 6- nitrine hexanoyl hydrazines（0.55 g, yield：56.2%）；
6- nitrine hexanoyl hydrazines are weighed respectively（102.0 mg, 0.60 mmol）, DOX × HCl（115.9 mg, 0.20 mmol）And nothing
Water Na2SO4（103 mg, 0.73 mmol）Add in the round-bottomed flask with condensation reflux unit, the absolute methanol with 30 mL is molten
Solution.3 drop glacial acetic acid are subsequently added, after stirring, lucifuge reacts 24 h in 65 °C of oil baths.After reaction terminates, cotton is used
Flower filtering, removes the Na in reaction solution2SO4, remove and precipitated three times in ice ether after solvent.Finally, obtained down using centrifugal process
The sediment in face, places it in and 24 h is dried in vacuum drying chamber, obtain the product of red powder, as N3-hyd-DOX
×HCl（93.5 mg, yield：62.4%）.
Embodiment two：3,3 '-two sulphur diine butyl dipropionates of alkynyl end-blocking（B-ss-B）Synthesis
First, with 3,3 '-dithiodipropionic acid and 3- butyne-1-ols for raw material, with 1- ethyls-（3- dimethylaminopropyls）Carbon
Diimmonium salt hydrochlorate is activator, and DMAP is catalyst, and dichloromethane is solvent, by esterification, is obtained
3,3 '-two sulphur diine butyl dipropionates.Specific synthetic method is as follows：
Glass apparatus used is placed in 120 °C of baking ovens and dries 4 h, taking-up is placed in desiccator that to be cooled to room temperature stand-by.It is first
First, 3,3 '-dithiodipropionic acid is sequentially added into three-neck flask（3.66 g, 17.40 mmol）, 1- ethyls-（3- dimethyl
Aminopropyl）Carbodiimide hydrochloride（EDC×HCl）（8.67 g, 45.23 mmol）And DMAP（DMAP）
（3.19 g, 2.61 mmol）, with 100 mL CH2Cl2Dissolving；Secondly, by 3- butyne-1-ols（3.17 g, 45.23 mmol）
With 15 mL CH2Cl2Add in constant pressure funnel.2 h are added dropwise in ice-water bath, after completion of dropwise addition, device is moved on to 25 °C
Oil bath in, continue react 48 h；
After reaction terminates, rotary evaporation removes a small amount of solvent, and filter paper is filtered to remove the salt of reaction generation, then with 150 mL's
CH2Cl2Dissolving, with 1 M HCl solutions and saturated sodium-chloride（NaCl）The mixed solution of the aqueous solution is extracted 2 times, the water layer of collection
With 30 mL CH2Cl2Extraction 3 times.After organic layer is collected, anhydrous Na is used2SO4Dry 4 h, remove solvent after crude product after
Continuous to carry out separating-purifying using column chromatography, eluent is the mixed solution of ethyl acetate and petroleum ether（Ethyl acetate：Petroleum ether
=1：6）.Final product is collected, and purer 3,3 '-two sulphur diine butyl dipropionate B- are obtained after 24 h of vacuum dryingss-
B（3.63 g, yield：59.4%）.
Embodiment three：The acetal radical polyethylene glycol of two azidoethyl two of azido end-blocking（N3-a-PEG21-a-N3）Synthesis
The acetal radical polyethylene glycol of two azidoethyl of compound two of azido end-blocking（N3-a-PEG21-a-N3）, pass through two-step reaction
Obtain：First, the acetal radical polyethylene glycol of Dichloroethyl two is synthesized（Cl-a-PEG21-a-Cl）；Secondly, two azidoethyls two are synthesized
Acetal radical polyethylene glycol（N3-a-PEG21-a-N3）.Specific synthetic method is as follows：
The first step, synthesizes the acetal radical polyethylene glycol of Dichloroethyl two（Cl-a-PEG21-a-Cl）.Take the poly- second that molecular weight is 1000
Glycol（HO-PEG22-OH）（6.01 g, 6.01 mmol）With p-methyl benzenesulfonic acid pyridine（PPTS）（0.30 g, 1.20 mmol）
In the side-neck flask for being placed in 100 mL, and 50 mL toluene is added, impurity is removed using the method for two sub-atmospheric pressure toluene azeotropic
Water.After after its cooling, under nitrogen protection, the CH that 30 mL are dried is added2Cl2Dissolving, and add in constant pressure funnel 10
ML CH2Cl2With 2- chloroethyl vinyl ethers（CEVE）（3.20 g, 30.05 mmol）, after ice-water bath is dripped, at room temperature after
1 h of continuous reaction, the mass fraction for adding 10 mL is 5% sodium carbonate（Na2CO3）Aqueous solution terminating reaction；
The method that the purification of product is combined using extraction and precipitated phase.First use CH2Cl2Dilute reaction solution, with 10 mL phosphoric acid buffers
Solution（PBS, pH 10.0) and the saturation NaCl aqueous solution mixed solution extraction, obtained water layer continues to use CH2Cl2Extraction three
It is secondary.Organic layer anhydrous Na2SO4Dry after 4 h, rotary evaporation removes CH2Cl2Solvent.Then, it is precipitated in n-hexane
Three times, collect sediment and be dried in vacuo, obtain product Cl-a-PEG21-a-Cl（5.75g, yield：78.9%）；
Second step, synthesizes the acetal radical polyethylene glycol of two azidoethyl two（N3-a-PEG21-a-N3）.The Cl- that upper step is obtaineda-
PEG-a-Cl（2.37 g, 1.95 mmol）And NaN3（0.76 g, 11.70 mmol）In the DMF for being dissolved in 25 mL, 60 °C of oil
48 h of the lower reaction of bath.After reaction terminates, neutral Al is used2O3Short column be filtered to remove unreacted NaN3, it is placed in 60 DEG C of oil baths,
DMF is removed under reduced pressure.Then, CH is used2Cl2Crude product is dissolved, with pH 10.0 PB cushioning liquid purification by liquid extraction products, the water of collection
20 mL of layer CH2Cl2Extraction.Most organic phase is collected at last, and uses anhydrous Na2SO4Dry, rotary evaporation removes solvent, very
Sky is dried to obtain the product N of brown3-a-PEG21-a-N3（1.68 g, yield：63.9%）.
Example IV：Polymer poly (the SS- of alkynyl end-blockingalt-A)nSynthesis
Under nitrogen protection, by 3,3 '-two sulphur diine butyl dipropionates（B-ss-B）With the poly- second of the acetal radical of two azidoethyl two
Glycol（N3-a-PEG21-a-N3）Chemically react, prepare the polymer poly (SS- of alkynyl end-blockingalt-A)n.Specific synthesis
Method is as follows：
Under the protection of nitrogen, CuBr is added into the side-neck flask of the DMF containing 10 mL（0.085 g, 0.59 mmol）With
Pentamethyl-diethylenetriamine（PMDETA）（0.102 g, 0.59 mmol）, stirring is complexed both, is then sequentially added
The B- of synthesisss-B（0.25 g, 0.79 mmol）And N3-a-PEG21-a-N3（0.97 g, 0.79 mmol）.Take out inflation three times
Afterwards, good seal is placed in 50 °C of oil baths and reacts 48 h device under nitrogen protection, then, adds 10 mol% B-ss- B continues
24 h are reacted, to ensure two ends as butynyl.Crude product is dialysed with DMF（Bag filter molecular cut off is 3500 Da）, often
The dialyzate renewed every a period of time.After 48 h, DMF is removed under reduced pressure in 60 °C of oil pumps.Then with 150 mL CH2Cl2Again it is molten
Solution, adds the saturation NaCl aqueous solution and removes mantoquita.Isolated organic layer anhydrous Na2SO4Dry after 4 h, rotary evaporation is removed
Solvent is removed, then is precipitated 3 times in diethyl ether solution, sediment is collected and is dried in vacuo, sticky product poly (SS- are finally givenalt-A)n（0.74 g, yield：60.7%）.
Embodiment five：The adriamycin prodrug DOX- of double-responsehyd-poly(SS-alt-A)n-hyd- DOX synthesis
Utilize the doxorubicin derivative and polymer poly (SS- of desalting processingalt-A)nChemically react, obtain reduction/pH
Dual responsiveness adriamycin prodrug DOX-hyd-poly(SS-alt-A)n-hyd-DOX.Specific synthetic method is as follows：
First, to doxorubicin derivative（N3-hyd-DOX×HCl）Carry out desalination acid treatment.In round-bottomed flask, N is added3-hyd-DOX×HCl（0.095 g, 0.13 mmol）, dissolved with 3 mL DMSO, and add 10 mL triethylamines（TEA）Stirring 2
Supernatant is siphoned away after h, standing and adds new TEA solution continuation stirring, is so repeated 2 times, finally leaves and takes DMSO layers of sample
It is stand-by.In side-neck flask, lead in the state of nitrogen, first add CuBr（0.015 g, 0.104 mmol）And PMDETA（0.018
G, 0.104 mmol）, dissolved with 5 mL DMF solvents and stir 5 min, then add poly (SS- into bottle successivelyalt-A)n
（0.23 g, 0.052 mmol）With the above-mentioned N for sloughing hydrochloride3-hyd- DOX solution, takes out after inflating three times, by bottle seal
It is placed in 50 °C of oil baths and reacts 48 h.After reaction terminates, reaction solution is moved on in the bag filter that molecular cut off is 5000 Da,
Dialysed using deionized water, every changing a deionized water after 4 h.Dialyse after 72 h, solution in bag filter is taken out,
It is placed in freeze dryer and freezes after being freezed in refrigerator, finally obtains Red viscous solid product DOX-hyd-poly(SS-alt-A)n-hyd-DOX（0.26 g, yield：84.2%）.
It is utilized respectively proton nmr spectra（1H NMR）And infrared spectrum（FT-IR）Verify gained micromolecular compound, gather
The structure of compound and reduction/pH dual responsiveness adriamycin prodrugs based on polyethylene glycol.Accompanying drawing 1 is doxorubicin derivative
（N3-hyd-DOX×HCl）Hydrogen nuclear magnetic resonance spectrogram, accompanying drawing 2 be the sulphur diine butyl dipropionate of polymer 3,3 '-two core
Magnetic resonance hydrogen spectrogram, accompanying drawing 3 are Cl-a-PEG21-a- Cl and N3-a-PEG21-a-N3Hydrogen nuclear magnetic resonance spectrogram, accompanying drawing 4 be poly-
Compound poly (SS-alt-A)3.6Hydrogen nuclear magnetic resonance spectrogram, accompanying drawing 5 be DOX-hyd-poly(SS-alt-A)3.6-hyd-DOX
Hydrogen nuclear magnetic resonance spectrogram, accompanying drawing 6 be HO-PEG21-OH, Cl-a-PEG21-a-Cl, N3-a-PEG21-a-N3, poly(SS-alt-A)3.6, DOX-hyd-N3And DOX-hyd-poly(SS-alt-A)3.6-hyd- DOX infrared spectrogram.With reference to accompanying drawing 1,
2nd, 3,4,5,6 can meet experimental design with proved response product.
Embodiment six：Polymeric prodrugs micella is prepared using dialysis
Take polymeric prodrugs（DOX-hyd-poly(SS-alt-A)3.6-hyd-DOX）25 mg are dissolved in 2 mL DMF, stirring
After 2 h, with micro syringe with 2 mL h-1Speed 20 mL deionized water is added dropwise into solution, after completion of dropwise addition, continue
Stir 2 h.Then, solution is fitted into the bag filter that molecular cut off is 5000 Da, will after 24 h of deionized water dialysis
Solution takes out and is settled to 25 mL, finally gives concentration for 1 mg mL-1Micellar solution.Accompanying drawing 7 is DOX-hyd-poly
(SS-alt-A)3.6-hydThe transmission electron microscope photo for the micella that-DOX is self-assembly of in water（A）With dynamic light scattering curve
（B）；Such as：Accompanying drawing 7（A）It show the pattern that polymeric prodrugs are self-assembly of spherical micelle in aqueous；Accompanying drawing 7（B）Institute
It is shown as the dynamic light scattering test curve figure of correspondence micella particle diameter, it can be seen that polymeric prodrugs micella average grain diameter is 48 nm.
Embodiment seven：The vitro drug release test of polymeric prodrugs
The micellar solution for taking 4 mL well prepared in advance（Micellar concentration is 1 mg mL-1）It is 7000 Da's to be placed in molecular cut off
In bag filter, two ends, which are sealed, to be placed in 50 mL big centrifuge tube, then, 30 mL is separately added into corresponding centrifuge tube different
Cushioning liquid.Cushioning liquid is divided into four kinds：（1）Phosphate buffer（The mM of pH 7.4,10）；（2）The phosphorus of GSH containing 10 mM
Acid buffer（The mM of pH 7.4,10）；（3）Acetate buffer solution（The mM of pH 5.0,10）；（4）The acetic acid of GSH containing 10 mM delays
Fliud flushing（The mM of pH 5.0,10）.Then, big centrifuge tube is placed in 37.5 °C of constant temperature oscillation instrument, entered with 160 r/min speed
Row vibration.At the time point of setting, 5 mL release liquid is taken out successively and corresponding cushioning liquid is added.Every group of experiment carries out 3
Individual balance test, finally averages.The release liquid of taking-up is measured with sepectrophotofluorometer to DOX concentration.Be loaded with Ah
Release profiles of the polymer micelle of mycin under the conditions of different reduction and pH as shown in Figure 8, pH 5.0 or containing
Have in 10 mM GSH phosphate buffer, drug release rate is significantly faster than that under normal physiological conditions；Meanwhile, containing 10
Medicine adds up release rate maximum in the mM GSH cushioning liquid of pH 5.0.Insoluble drug release illustrates that the polymeric prodrugs micella has
Obvious reduction and pH dual responsiveness, can reach the purpose of control release.
Embodiment eight：Polymeric prodrugs micella suppresses tumor cell proliferation performance test
Respectively by human cervical carcinoma cell (HeLa cells) and human liver cancer cell（HepG2 cells）Culture is being supplemented with 10%
In the DMEM culture mediums of hyclone (FBS), 37 DEG C, 5% CO are placed in2Cultivated in the incubator of (relative humidity is 90%), periodically
Change nutrient solution.Selection is in the cell of active growth phase and is inoculated in 96 orifice plates that 100 μ L DMEM culture mediums are contained in every hole,
Cultivate 24 h.With dialysis configuration polymeric prodrugs DOX-hyd-poly(SS-alt-A)3.6-hyd- DOX micella mother liquor（Ah
Mycin concentration is 94 mg L-1）, a series of micellar solution of various concentrations is added in 96 orifice plates, continues to cultivate 48 h respectively
With 72 h.25 μ L MTT reagents are subsequently added into, further after 4 h of culture, are existed with ELIASA (Bio-Rad model 680)
Corresponding absorbance is measured under 570 nm.The computational methods of cell survival rate are：Cell survival rate (Cell viability)
(%) = [A]test/[A]control× 100%, wherein, [A]testFor DOX-hyd-poly(SS-alt-A)3.6-hyd- DOX gathers
The absorbance measured in the presence of compound prodrug micelle, and [A]controlFor the extinction measured in the case of without polymeric prodrugs
Degree.Each sample test three times, takes its average value.As shown in Figure 9,（A）With（B）Respectively with the h of HeLa cell culture 48 and
The survival rate of cell after 72 h；（C）With（D）Respectively with the survival rate of cell after the h of HepG2 cell culture 48 and 72 h.As a result
Display：Compared with anticancer drugs, doxorubicin, synthesized polymeric prodrugs have good suppression HeLa cells and HepG2 cells
The ability of propagation.
The method comprises the steps of firstly, preparing doxorubicin derivative（N3-hyd-DOX×HCl）；Prepare small point of alkynyl end-blocking respectively again
The sub- sulphur diine butyl dipropionate of compound 3,3 '-two（B-ss-B）The acetal of two azidoethyl of compound two blocked with azido
Base polyethylene glycol（N3-a-PEG-a-N3）；Then, the polymer poly containing acetal bonds and disulfide bond is prepared by polymerisation
(SS-alt-A)；Finally, then by reacting adriamycin is connected to polymer two ends, obtained with reduction and pH double-responses
Adriamycin prodrug DOX-hyd-poly(SS-alt-A)3.6-hyd- DOX, changes other catalyst and part that the present invention is limited
Product can be equally obtained, the material rate that the molecular weight and the present invention of polyethylene glycol are limited is changed, different m values can be obtained
The product of n values；What the present invention was limited there is stimulating responsive polymer prodrug can be assembled into prodrug micelle simultaneously in aqueous
It is stabilized, and under acid either reducing condition, sensitive groups or segment can occur fracture and be destroyed micella, so that
The medicine that can be will accumulate in inside micella is rapidly discharged, for treating, and achieves the technology effect recorded such as embodiment
Really.
Claims (10)
1. a kind of reduction/pH dual responsiveness adriamycin prodrugs, it is characterised in that expressed by the following chemical structure formula：
In formula, m is that 3~113, n is 3~15.
2. the preparation method of reduction described in claim 1/pH dual responsiveness adriamycin prodrugs, it is characterised in that including following
Step：
(1) using 6- nitrine hexanoyl hydrazines and doxorubicin hydrochloride as raw material, glacial acetic acid is catalyst, and Ah mould is obtained by reacting
Plain derivative；
(2) under the conditions of inert gas atmosphere, in the presence of copper salt catalyst and part, with 3,3 '-two sulphur diine butyl dipropyls
Acid esters and the acetal radical polyethylene glycol of two azidoethyl two are raw material, and the polyethylene glycol alternating that two end alkynyl radicals are blocked is prepared by reacting
Copolymer；
(3) in the presence of copper salt catalyst and part, the polyethylene glycol that two end alkynyl radicals obtained with step (2) are blocked replaces
Copolymer reacts with the doxorubicin derivative that step (1) is obtained, and prepares before the reduction/pH dual responsiveness adriamycins
Medicine.
3. the preparation method of reduction/pH dual responsiveness adriamycin prodrugs according to claim 2, it is characterised in that step
(1) in, 6- bromocaproic acids methyl esters and reaction of sodium azide obtain 6- nitrine methyl caproates, then 6- nitrine methyl caproate and hydration
Hydrazine reaction, obtains 6- nitrine hexanoyl hydrazines；It is raw material, 1- using 3,3 '-dithiodipropionic acid and 3- butyne-1-ols in step (2)
Ethyl-（3- dimethylaminopropyls）Carbodiimide hydrochloride is that activator, DMAP are catalyst, passes through esterification
Reaction, obtains 3,3 '-two sulphur diine butyl dipropionates；Polyethylene glycol is used to be urged with 2- chloroethyl vinyl ethers for raw material, acid
In the presence of agent, reaction obtains the polyethylene glycol product that end is chloro, and then end is the polyethylene glycol product of chloro with folding
Sodium nitride reaction obtains the acetal radical polyethylene glycol of two azidoethyl two；In step (3), it will be used after doxorubicin derivative desalting processing
In reaction.
4. the preparation method of reduction/pH dual responsiveness adriamycin prodrugs according to claim 2, it is characterised in that：Step
(1) in, the temperature of reaction is 60 DEG C~80 DEG C, and time of reaction is the h of 12 h~24；In step (2), inert gas is nitrogen；
The temperature of reaction is 25 DEG C~50 DEG C, and time of reaction is the h of 24 h~72；In step (3), the temperature of reaction is 25 DEG C~50
DEG C, time of reaction is the h of 24 h~48.
5. the preparation method of reduction/pH dual responsiveness adriamycin prodrugs according to claim 2, it is characterised in that：Mantoquita
Catalyst is selected from Salzburg vitriol, stannous chloride or cuprous bromide；Part is selected from sodium ascorbate, bipyridyl, pentamethyl two
One kind in ethylenetriamine, tetramethylethylenediamine or hexamethyl trien.
6. the preparation method of reduction/pH dual responsiveness adriamycin prodrugs according to claim 2, it is characterised in that：Step
(1) in, the mol ratio of 6- nitrine hexanoyl hydrazines and doxorubicin hydrochloride is 1：(2~6)；In step (2), the acetal of two azidoethyl two
The mol ratio of base polyethylene glycol, 3,3 '-two sulphur diine butyl dipropionates and copper salt catalyst is 1: (1.1~1.5):（0.5~
1.5）；Copper salt catalyst and the mol ratio of part are 1: (1~2)；In step (3), the polyethylene glycol of two end alkynyl radicals end-blocking is handed over
It is 1 for the mol ratio of copolymer, doxorubicin derivative：(2~5).
7. the preparation method of reduction/pH dual responsiveness adriamycin prodrugs according to claim 3, it is characterised in that：Often walk
After reaction terminates, all by purification processes.
8. a kind of preparation method of prodrug micelle, it is characterised in that comprise the following steps：
(1) using 6- nitrine hexanoyl hydrazines and doxorubicin hydrochloride as raw material, glacial acetic acid is catalyst, and Ah mould is obtained by reacting
Plain derivative；
(2) under the conditions of inert gas atmosphere, in the presence of copper salt catalyst and part, with 3,3 '-two sulphur diine butyl dipropyls
Acid esters and the acetal radical polyethylene glycol of two azidoethyl two are raw material, and the polyethylene glycol that two end alkynyl radicals are blocked is prepared by reaction
Alternate copolymer；
(3) in the presence of copper salt catalyst and part, the polyethylene glycol that two end alkynyl radicals obtained with step (2) are blocked is handed over
The doxorubicin derivative obtained for copolymer and step (1) reacts, and prepares reduction/pH dual responsiveness adriamycin prodrugs；
(4) reduction/pH dual responsiveness adriamycin prodrugs are self-assembly of prodrug micelle in aqueous.
9. a kind of preparation method of stimulating responsive anticancer nano drug system, it is characterised in that comprise the following steps：
(1) using 6- nitrine hexanoyl hydrazines and doxorubicin hydrochloride as raw material, glacial acetic acid is catalyst, and Ah mould is obtained by reacting
Plain derivative；
(2) under the conditions of inert gas atmosphere, in the presence of copper salt catalyst and part, with 3,3 '-two sulphur diine butyl dipropyls
Acid esters and the acetal radical polyethylene glycol of two azidoethyl two are raw material, and the polyethylene glycol that two end alkynyl radicals are blocked is prepared by reaction
Alternate copolymer；
(3) in the presence of copper salt catalyst and part, the polyethylene glycol that two end alkynyl radicals obtained with step (2) are blocked is handed over
The doxorubicin derivative obtained for copolymer and step (1) reacts, and prepares reduction/pH dual responsiveness adriamycin prodrugs；
(4) reduction/pH dual responsiveness adriamycin prodrugs are self-assembly of prodrug micelle in aqueous；
(5) prodrug micelle and buffer solution are mixed to get stimulating responsive anticancer nano drug system.
10. reduction described in claim 1/pH dual responsiveness adriamycin prodrugs are in stimulating responsive anticancer nano medicine is prepared
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| CN107641201A (en) * | 2017-11-01 | 2018-01-30 | 苏州大学 | Block copolymer preparation method and applications of the Quick Oxidation/reduction dual responsiveness containing double selenium keys |
| CN107641201B (en) * | 2017-11-01 | 2020-07-17 | 苏州大学 | Preparation method and application of block copolymer containing double selenium bonds with rapid oxidation/reduction dual responsiveness |
| CN108283720A (en) * | 2018-01-22 | 2018-07-17 | 苏州大学 | It is bonded the polyphosphate prodrug and the preparation method and application thereof of camptothecine and adriamycin simultaneously |
| CN112940248A (en) * | 2021-02-04 | 2021-06-11 | 西安交通大学 | PH-responsive metal coordination poly-prodrug nanoparticles and preparation method thereof |
| CN112940248B (en) * | 2021-02-04 | 2023-07-18 | 西安交通大学 | PH responsive metal coordination polymer prodrug nano-particle and preparation method thereof |
| CN113912841A (en) * | 2021-10-22 | 2022-01-11 | 清华大学深圳国际研究生院 | PH and Redox dual-response diblock amphiphilic polymer prodrug and preparation method thereof |
| CN114644665A (en) * | 2022-04-20 | 2022-06-21 | 湖南工程学院 | light/pH dual-response coupled prodrug compound and preparation method and application thereof |
| CN114644665B (en) * | 2022-04-20 | 2023-05-12 | 湖南工程学院 | photo/pH dual-response coupled prodrug compound and preparation method and application thereof |
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| Publication number | Publication date |
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| CN110591078B (en) | 2021-11-05 |
| CN110591078A (en) | 2019-12-20 |
| CN107141323B (en) | 2019-10-29 |
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